Mesenchymal stem/stromal cells (MSCs) hold great promise in regenerative medicine due to their ability to modulate immune responses and regenerate damaged tissues, particularly in cartilage repair. Cryopreservation is essential for their long-term storage and future therapeutic use. However, the effects of prolonged cryopreservation—especially when cells are stored for several years in liquid nitrogen—on their viability and functional potential remain poorly understood.
This study aimed to assess whether extended cryopreservation impacts the viability and colony-forming ability of synovium-derived primary MSCs isolated from post-mortem donors. A total of 22 primary cell samples (21 synovium-derived and one bone marrow-derived, used as a reference) were thawed after being cryopreserved for periods ranging from two to over five years. Following thawing and in vitro culture, the colony-forming unit fibroblast (CFU-F) assay was used to evaluate their proliferative potential.
Out of 22 samples, only five successfully formed colonies. Statistical analysis revealed no significant correlation between storage duration and colony-forming capacity (Spearman's r = –0.1225, p = 0.5528). Similarly, no statistically significant difference in storage times was found between successfully and unsuccessfully cryopreserved samples (p = 0.157, Mann-Whitney U test).
While the low recovery rate suggests that prolonged cryopreservation may negatively affect MSC viability, the findings indicate that storage time alone is not a determining factor. Instead, donor variability, post-mortem tissue conditions, and handling procedures likely play a more critical role in cryopreservation outcomes.
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